Methods Improving HIV Protein Expression: Cell Substrate and Protein Purification: Despite the widespread use of GMP-established pharma cell substrates (e.g., CHOs, 293 etc.) in development of recombinant HIV Env protein antigens, critical bottlenecks still exist in their use for large-scale, high-yield GMP manufacturing; yields often are on the order of mg/L compared to mAbs at gm/L. Some of the limitations relate to their intrinsic incapacities to metabolically produce high levels of stable properly folded, properly glycosylated recombinant HIV Env protein, often times requiring extensive clonal screening to identify the rare high-level producer clone. These constraints have a cascading effect in increasing the overall cost and time for production of HIV vaccine antigens from millions of dollars and years of upstream and downstream process development. The objective is to evaluate and modulate the molecular pathways involved in regulating and enhancing HIV envelope/antigen expression in mammalian cell lines and to accelerate development of purification platforms in a CGMP manufacturing setting.